GROWTH FACTOR RECEPTOR SIGNAL TRANSDUCTION

这项研究计划的基础是， 扩大我们目前资助的计划项目，该项目已成功地 综合互补的科学推力来研究生长因子 受体介导的信号转导机制在正常和 恶性细胞 在这一计划中，七个精心 在四个核心设施的支持下， 利用各种模型系统来测试假设， 在质膜，细胞质， 核水平。 项目1（R. Davis）和2（H.罗宾逊和 D. Gamett）将寻求确定受调节的磷酸化位点 对EGF受体和erB B蛋白的影响，并评估其作用 调节受体功能和ER B B疾病潜能。 项目3（M. Czech and J. Klarlund）将检验以下假设： 多亚基丝氨酸激酶酪蛋白的酪氨酸磷酸化 激酶II，激活后一种酶，作为主要的 受体和癌基因信号磷酸化级联反应 酪氨酸激酶 项目4（G.约翰逊）试图定义 生长因子受体中特异性G蛋白的功能 使用体外诱变和cDNA表达的信号传导事件 对Gi alpha蛋白质来说。 本研究将评估 与其他G蛋白亚基一起表达α亚基， 在调节对生长因子的代谢反应中的作用。 项目 5（F. Fay）将探索生长因子- 介导的趋化反应，通过分析局部变化， 细胞内钙和（H+）使用基于计算机图像 中性粒细胞中荧光信号的增强。 项目 6（J. Massague）和7（J. and G.和G. Stein）涉及的努力 这两个小组致力于定义TGF-β作用的分子基础。 项目6将重点关注TGF-β调节 细胞外膜受体的表达和功能 基质分子（整合素）。 项目7将寻求确定 特异性基因，其在存在下差异表达， 和缺乏TGF-β，并参与调节3 T3- L1细胞分化。 四个核心设施将提供 上述项目的支持、仪器、技术和试剂 综合努力，提供（1.）媒体和技术 组织培养时间，（2.）肽合成，（3.）细胞科学 包括免疫组织学和显微操作的技术， （4.）重组DNA技术和试剂。 核心设施将 由经验丰富的资深调查员指导 以及每个核心提供的专门知识领域的培训。

This proposed research program builds upon and significantly expands our currently funded Program Project which has successfully integrated complementary scientific thrusts to study growth factor receptor-mediated signal transduction mechanisms in normal and malignant cells. In this proposed Program, seven carefully integrated projects with the support of four core facilities will utilize a variety of model systems to test hypotheses about cellular signalling pathways at the plasma membrane, cytoplasmic, and nuclear levels. Projects 1 (R. Davis) and 2 (H. Robinson and D. Gamett) will seek to identify regulated phosphorylation sites on the EGF receptor and erb B proteins and to evaluate their role in modulating receptor function and erb B disease potential. Project 3 (M. Czech and J. Klarlund) will test the hypothesis that tyrosine phosphorylation of a multi-subunit serine kinase, casein kinase II, activates this latter enzyme as part of a major signalling phosphorylation cascade for receptor and oncogene tyrosine kinases. Project 4 (G. Johnson) seeks to define the function of specific G proteins in growth factor receptor signalling events using in vitro mutagenesis and expression of cDNA for Gi alpha proteins. This study will assess interactions of expressed alpha subunits with other G protein subunits and their role in mediating metabolic responses to growth factors. Project 5 (F. Fay) will explore mechanisms that underly growth factor- mediated chemotactic responses by analysis of local changes in intracellular calcium and (H+) using computer-based image intensification of fluorescence signals in neutrophils. Projects 6 (J. Massague) and 7 (J. and G. and G. Stein) involve efforts of two groups toward defining the molecular basis of TGF-beta action. Project 6 will focus on the mechanisms by which TGF-beta modulate expression and function of cell surface receptors for extracellular matrix molecules (integrins). Project 7 will seek to identify specific genes which are differentially expressed in the presence and absence to TGF-beta and which are involved in regulating 3T3- L1 cell differentiation. Four core facilities will provide support, instrumentation, technology and reagents for the above integrated efforts by making available (1.) media and technical time for tissue culture, (2.) peptide synthesis, (3.) cell science technology including immunohistology and micromanipulations and, (4.) recombinant DNA techniques and reagents. Core facilities will be directed by established investigators with extensive experience and training in the areas of expertise provided by each core.